Catielen Paula Pavi

Milk and dairy products may be contaminated by enteric viruses along the production chain, and their high protein and fat content hinders viral concentration and molecular detection. Internationally standardized recovery protocols have been established for leafy greens, soft fruits, and bivalve mollusks, but remain unavailable for dairy matrices. Three low-cost methods were compared for pasteurized milk, namely polyethylene glycol precipitation (PEG), Tris-Glycine-Beef Extract elution (TGBE), and a modified organic flocculation (FLO) exploiting the matrix’s own casein, and three for mozzarella cheese (PEG, TGBE, and glycine buffer–chloroform extraction, GBC). Samples were spiked with murine norovirus 1 (MNV-1) and murine hepatitis virus 3 (MHV-3) as non-enveloped and enveloped surrogates, and recovery was determined by RT-qPCR. TGBE yielded the highest mean recovery in both matrices (milk: 1.5 ± 0.6% for MNV-1 and 4.3 ± 0.6% for MHV-3; cheese: 4.7 ± 3.5% and 15.1 ± 17.5%), and its superiority over PEG for MNV-1 in milk was the only difference to reach statistical significance. PEG achieved recovery comparable to that of TGBE for MNV-1 in cheese (1.1 ± 1.5%) but with greater variability, whereas GBC recovery was negligible and the protocol requires a hazardous solvent. Organic flocculation yielded lower recovery (0.28 ± 0.34% for MNV-1) but allowed for processing of one-liter volumes, a potential alternative for large-volume screening of bulk milk. TGBE therefore appears the most practical candidate for viral recovery from dairy matrices in laboratories with limited infrastructure, although a larger number of replicates and validation with naturally contaminated samples remain necessary before routine application.

Meat products represent important vehicles for foodborne viral transmission, yet standardized detection methodologies for these complex matrices remain limited. This study systematically compared three viral recovery methods across nine meat products representing chicken, pork, and processed deli meat matrices: polyethylene glycol 6000 (PEG) precipitation, glycine buffer-chloroform (GBC) phase separation, and β-mercaptoethanol (B-M) chaotropic lysis. Using Murine Norovirus 1 (MNV-1) and Murine Hepatitis Virus 3 (MHV-3) as process controls, recovery efficiency was assessed by RT-qPCR. The B-M method demonstrated the most consistent performance, successfully detecting MNV-1 across all tested matrices and significantly outperforming PEG and GBC for MNV-1 in chicken and pork. Applying the B-M method to 25 commercial retail products revealed the genetic material of Hepatitis E virus (HEV) in four samples and Porcine Circovirus type 2 (PCV-2) in one sample, exclusively within pork products. These findings provide critical comparative performance data for method selection in viral surveillance of meat and document the occurrence of HEV and PCV-2 genetic material in commercial pork products from Santa Catarina, Brazil, highlighting a significant food and agricultural biosafety concern in this major meat-producing and exporting state.

As interações entre vírus entéricos e micropoluentes ambientais, como fármacos, pesticidas e microplásticos, podem afetar a sobrevivência, disseminação e infectividade viral, representando um desafio emergente para a saúde pública e ambiental. Fatores físicos e químicos, como pH, salinidade e matéria orgânica, modulam a adsorção e a estabilidade viral, ampliando riscos de contaminação hídrica e alimentar. Pesticidas e fármacos podem induzir estresse oxidativo, comprometendo defesas celulares e favorecendo infecções. Entre os micropoluentes, os microplásticos se destacam pela persistência e pela formação de biofilmes, capazes de prolongar a estabilidade viral. Essas interações, entretanto, podem tanto proteger quanto reduzir a infectividade, dependendo das condições ambientais e do vírus envolvido. Persistem lacunas metodológicas no monitoramento simultâneo de vírus e micropoluentes, dada a necessidade de abordagens analíticas distintas e a baixa eficiência dos sistemas convencionais de tratamento na remoção desses contaminantes. Assim, são necessárias estratégias integradas que unam pesquisa aplicada, monitoramento ambiental e políticas públicas para reduzir riscos ao ambiente e à saúde humana.

Emerging pollutants encompass a diverse array of chemicals classified as micropollutants, including pharmaceuticals and microplastics (MPs). Viruses are known to adsorb onto MPs, including binding to bacterial biofilms that form the plastisphere. In this review, we conducted an extensive bibliographic survey to critically assess the potential interactions between micropollutants, such as MPs, organic micropollutants and viruses in aquatic environments, within the One Health context. The interaction between viruses and bacterial cell wall components can increase the infectivity and thermal stability of viral particles, which thrive on biofilms commonly found on MPs in aquatic systems. MPs, acting as viral vectors, can impact viral life cycles, survival, transmission and interaction with hosts, posing significant risks to human, animal and environmental health. There is a clear need for additional practical studies to understand how viruses remain stable when in contact with micropollutants. This field of research provides opportunities to better understand the broader impacts of these interactions, including the potential for new viral outbreaks due to the prolonged persistence of pathogens in the environment.

Noroviruses are the leading cause of gastroenteritis outbreaks in humans worldwide. Their unique properties ensure stability over extended periods under adverse conditions, which enhances their risk as food and water contaminants. In recent years, intensive research has focused on the natural antimicrobial potential of plant metabolites as disinfectants against environmental pathogens. The oregano essential oil (OEO) has gained attention due to its valuable properties, including antimicrobial, antioxidant, antiviral, and antifungal activities. However, the susceptibility of OEO to degradation and oxidation under environmental or storage conditions, coupled with its low water solubility, has limited its practical applications. Nanoencapsulation has emerged as a promising strategy to overcome these limitations by prolonging shelf life, improving stability, enabling controlled release, and expanding its potential uses. In this study, we evaluated the virucidal potential of chitosan-based polymeric nanoparticles incorporating Origanum vulgare essential oil against murine norovirus 1 (MNV-1) for food and environmental applications. To assess the virucidal effect of the OEO nanoparticles, the reduction in viral infectivity was determined by comparing the TCID₅₀/mL values of untreated viral suspensions with those treated with the tested compounds at varying concentrations. The results demonstrated effective viral inactivation at all tested concentrations, with the undiluted formulation (40 mg/mL incorporated OEO) achieving the highest inactivation rate (99.72%). The blank formulation showed no significant virucidal activity, while the pure OEO exhibited cytotoxicity at most tested concentrations. These findings support the development of a biotechnological disinfectant with potential applications in both environmental and controlled conditions.

The Hepatitis E virus (HEV) is responsible for causing Hepatitis E, a zoonotic disease that has emerged as a significant global health concern, accounting for about 20 million infections and 70,000 deaths annually. Although it is often recognized as a disease that is acute in low-income countries, HEV has also been recognized as a zoonotic disease in high-income countries. The zoonotic transmission requires flexible approaches to effectively monitor the virus, vectors, and reservoirs. However, the environmental monitoring of HEV presents additional challenges due to limitations in current detection methods, making it difficult to accurately assess the global prevalence of the virus. These challenges hinder efforts to fully understand the scope of the disease and to implement effective control measures. This review will explore these and other critical concerns, addressing gaps in HEV research and highlighting the need for improved strategies in the monitoring, prevention, and management of Hepatitis E using a One Health approach.

Chikungunya virus (CHIKV), the causative agent of the chikungunya fever, is an alphavirus widely transmitted by the bite of the female mosquito of the genus Aedes sp., especially in tropical and subtropical regions. Brazil is the country most affected by the microorganism. CHIKV classically induces articular pain, which can become long lasting for even years in a great number of the infected individuals, reducing their quality of life. The mechanisms of CHIKV-induced pain are poorly understood, but recent evidence indicated a role for the transient receptor potential vanilloid 1 (TRPV1) in this pathology. Herein, we assessed the ability of intra-articularly injected inactivated CHIKV or its RNA to trigger nociception in mice. Both stimuli induced bilateral secondary hyperalgesia to mechanical and heat stimuli. These responses were attenuated by TRPV1 ablation or antagonism. Joint structural alterations and increased cartilage TRPV1 protein expression were detected in the ipsilateral knee joints injected with either CHIKV or viral RNA. However, the lack of this receptor did not influence the histological changes triggered by CHIKV or RNA. The results further support the role of TRPV1 in CHIKV-induced pain and highlight its importance in the chronic phase of the disease.

Os bacteriófagos, vírus que infectam exclusivamente bactérias, foram descobertos de forma independente por Frederick Twort (1915) e Félix d’Hérelle (1917). Esses vírus, parasitas intracelulares obrigatórios, possuem uma estrutura com capsídeo proteico que abriga material genético, podendo ser de DNA ou RNA. Sua abundância nos ecossistemas é notável, sendo responsáveis pelo controle populacional bacteriano, o que revela sua importância ecológica. Os fagos apresentam diferentes ciclos de replicação, com destaque para o ciclo lítico, devido à sua capacidade de destruir bactérias. A interação entre o fago e o hospedeiro ocorre por meio de proteínas específicas na superfície bacteriana, o que determina a especificidade de cada fago. A biotecnologia tem explorado cada vez mais os fagos, especialmente na terapia fágica, como uma alternativa aos antibióticos, oferecendo uma abordagem mais seletiva e com menos impacto sobre o microbioma do hospedeiro. A especificidade dos fagos permite agir contra patógenos sem afetar as bactérias benéficas. Além disso, técnicas como o encapsulamento de fagos têm sido utilizadas para melhorar sua estabilidade e eficácia, permitindo liberação controlada. Na agricultura, os fagos se apresentam como uma alternativa sustentável aos bactericidas convencionais, permitindo o controle de fitopatógenos sem induzir resistência bacteriana. Fagos também são usados no controle biológico de insetos, como pulgões e moscas. Na indústria alimentícia, os fagos têm sido empregados para combater patógenos como Salmonella e Escherichia coli, além de prolongar a vida útil dos alimentos, tornando-se uma ferramenta versátil e inovadora em diversas indústrias.

Os bacteriófagos, inicialmente explorados nas décadas de 1920 e 1930, caíram em desuso após a ascensão dos antibióticos, mas ressurgiram nas décadas seguintes devido ao aumento da resistência bacteriana. Eles atuam como controladores das populações bacterianas e podem transferir genes que aumentam a virulência, promovendo a diversidade genética das bactérias. Os fagos têm se mostrado uma ferramenta promissora no controle de doenças bacterianas em plantas, pecuária, aquicultura e alimentos, sendo eficazes contra patógenos como Salmonella, Escherichia coli e Listeria. Na agricultura, os fagos são usados para controlar fitopatógenos como Xanthomonas e Pseudomonas, substituindo biocidas convencionais. Na pecuária, ajudam a prevenir e tratar infecções bacterianas em animais de produção. Na aquicultura, fagos são aplicados contra patógenos marinhos, melhorando a saúde dos peixes e reduzindo a resistência a antibióticos. Em alimentos, são utilizados para controlar patógenos em superfícies de processamento e diretamente nos produtos. Exemplos incluem a redução de Escherichia coli na produção de queijo e a eliminação de Salmonella em carnes. Além disso, biofilmes de embalagens antimicrobianas com fagos aumentam a vida útil dos alimentos ao controlar o crescimento bacteriano. Os bacteriófagos também têm sido aplicados como biossensores para detectar patógenos em alimentos, oferecendo uma alternativa rápida e eficaz aos métodos tradicionais de cultura bacteriana. Diversos biossensores baseados em fagos foram desenvolvidos para detectar Salmonella, Listeria e outros patógenos, aumentando a sensibilidade e eficiência do diagnóstico.